CD4 + T淋巴细胞艾滋病毒的捕获使用一次性微流体芯片

Protocol

准备与抗CD4 + biotinlayted表面微流体通道。 10毫升的样品血液微流体通道注射泵，注射5毫升/分钟。 在20毫升/分钟为20分钟，用PBS冲洗出来的血。 15分钟，在5ul/min注入稀释的流式细胞仪，裂解液（从股票，用PBS液稀释至10％）。 孵育15分钟。 注射5毫升/分钟，20分钟主要的CD3 +抗体解决方案（从股票，用PBS液稀释至10％） 。 孵育2小时。 在5毫升/分…

Materials

Material Name	Type	Company	Catalogue Number	Comment
PDMS microchannel				Fabricated by SU-8 mold
Blood				Donated blood from normal human subject. Mass General Hospital, 10 X high density WBC or Brigham and Women’s Hospital.
PBS	Buffer	Invitrogen	14190	1x Dulbecco’s Phosphated Buffered Saline.
FACS lysing solution		Becton Dickinson	349202	Diluted to 10% from stock solution with PBS.
Anti-CD4+ biotinlayted Ab		Beckman Coulter	IM0704	Surface prepared with Anti-CD4+ biotinlayted Ab, diluted to 10% from stock solution with PBS/BSA after mixing with 0.2mg stock and 1ml deionized water.
goat anti-hCD3delta	pAB	Santa Cruz Biotechnology	sc-1128	Diluted to 10% from stock solution with PBS
rabbit anti-goat IgG PE		Santa Cruz Biotechnology	sc-3755	Secondary Ab fro anti-CD3 Ab, diluted to 10% from stock solution with PBS
anti-human CD4+		Becton Dickinson	557695	Alexa Fluor 488 labeled, diluted to 20% from stock solution with PBS.
DAPI		Invitrogen	D1306	4’,6-dianidino-2-phenylindole, dihydrochloride solution, diluted to 0.2% from stock solution with PBS.